ELISA/ BSA in capture antibody storage buffer - (Jul/10/2007 )

Hi all,

I have a question about ELISA. The antibodies (commercial) that I will be using as capture come in a buffer congaing ~ 2% BSA. Do I have to purify my Ab to get rid of BSA before immobilizing the antibodies on polystyrene plates? If so, what is the best way to do it?

Thank you in advance

Anna

I'm guessing your antibodies are going to be concentrated and you will have to dilute them significantly (100-fold or more) in your coating buffer/PBS before coating the plate. If this is the case I don't think it should matter.

Ceri

yah, what ceri told is quite correct

2% BSA is a SUBSTANTIAL amount of bsa to be in your ab!!! 20 mg/ml!!! What is the concentration of the ab? Your supplier should be able to provide you with ab that is protein free. At this concentration you will have significant competition between your ab and BSA sticking to the plate. For coating I have used ab at 50-100 ug/ml with 100-200 ul/well.

Even if you dilute your stock 100 fold your BSA is at 200 ug/ml!!!